| Summary: | This study presents a green protocol for the fabrication of a multifunctional smart nanobiocomposite (NBC) (ZnO-PIACSB-TiO<sub>2</sub>) for secure antimicrobial and antibiofilm applications. First, shrimp shells were upgraded to a polyimidazolium amphiphilic chitosan Schiff base (PIACSB) through a series of physicochemical processes. After that, the PIACSB was used as an encapsulating and coating agent to manufacture a hybrid NBC in situ by co-encapsulating ZnONPs and TiO<sub>2</sub>NPs. The physicochemical and visual characteristics of the new NBC were investigated by spectral, microscopic, electrical, and thermal methods. The antimicrobial indices revealed that the newly synthesized, PIACSB-coated TiO<sub>2</sub>–ZnO nanocomposite is an exciting antibiotic due to its amazing antimicrobial activity (MIC/MBC→0.34/0.68 μg/mL, 0.20/0.40 μg/mL, and 0.15/0.30 μg/mL working against <i>S. aureus</i>, <i>E. coli</i>, and <i>P. aeruginosa</i>, respectively) and antifungal capabilities. Additionally, ZnO-PIACSB-TiO2 is a potential fighter of bacterial biofilms, with the results being superior to those of the positive control (Cipro), which worked against <i>S. aureus</i> (only 8.7% ± 1.9 biofilm growth), <i>E. coli</i> (only 1.4% ± 1.1 biofilm growth), and <i>P. aeruginosa</i> (only 0.85% ± 1.3 biofilm growth). Meanwhile, the NBC exhibits excellent biocompatibility, as evidenced by its IC<sub>50</sub> values against both L929 and HSF (135 and 143 µg/mL), which are significantly higher than those of the MIC doses (0.24–24.85 µg/mL) that work against all tested microbes, as well as the uncoated nanocomposite (IC<sub>50</sub> = 19.36 ± 2.04 and 23.48 ± 1.56 µg/mL). These findings imply that the new PIACSB-coated nanocomposite film may offer promising multifunctional food packaging additives to address the customer demand for safe, eco-friendly food products with outstanding antimicrobial and antibiofilm capabilities.
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